Combination therapy for treatment of myeloproliferative neoplasms

ABSTRACT

The present application relates to treatment of myeloproliferative neoplasms using the JAK1/JAK2 inhibitor, ruxolitinib, in combination with a BET protein inhibitor, 2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one, wherein the combination is unexpectedly synergistic.

PRIORITY CLAIM

This application claims priority to U.S. Provisional Patent ApplicationNo. 63/034,214, filed on Jun. 3, 2020, and U.S. Provisional PatentApplication No. 63/078,123, filed on Sep. 14, 2020, each of which theentirety of which is incorporated by reference herein.

TECHNICAL FIELD

The present application relates to treatment of myeloproliferativeneoplasms using the JAK1/JAK2 inhibitor, ruxolitinib, in combinationwith a BET protein inhibitor,2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,wherein the combination is unexpectedly synergistic.

BACKGROUND

Compound 1 is a small molecule inhibitor of the acyl bindingbromodomains found within Bromodomain and Extraterminal Domain (BET)proteins. The four family members (BRD2, 3, 4, T) facilitatetranscription by binding acetylated histones and recruitingtranscription initiation and elongation complexes, thereby acting as alink between chromatin and transcriptional activation (see Shi, J., andVakoc, C. R., “The Mechanisms Behind the Therapeutic Activity of BETBromodomain Inhibition” Molecular Cell 2014; 54, 728-736, which isincorporated herein by reference in its entirety).

The BRD4 (and occasionally BRD3) gene is a component of a chromosomaltranslocation which yields an aggressive subtype of squamous carcinomastermed “NUT midline” carcinoma. These midline carcinomas are defined byt(15;19) translocations which give rise to the BRD4-NUT oncogene. Theresultant fusion protein consists of BRD4 at the N-terminus and NUT(nuclear protein in testis) at the C-terminus. Removal of this proteinfrom chromatin, either by RNA silencing or small molecule inhibitors,results in differentiation and growth arrest in these carcinomas (seeFilippakopoulos, P., et al., Selective Inhibition of BET Bromodomains.Nature 2010; 468, 1067-1073, which is incorporated herein by referencein its entirety).

The BET proteins are thought to exert proliferative effects byfacilitating transcription of oncogenes such as c-myc among others. BRD4inhibiting compounds have been shown to prevent BRD4 from associatingwith chromatin, including on the c-myc promoter, in multiple myelomamodels. This dissociation leads to decreased levels of c-myc expressionand decreased cell viability (see Delmore, J. E., et al., “BETBromodomain Inhibition as a Therapeutic Strategy to Target c-MYC” Cell2011; 146, 904-917, which is incorporated herein by reference in itsentirety). BRD4 has recently been discovered to bind to“super-enhancers” which are vast transcriptional regulatory elementswithin genes necessary for cell fate and survival. An example is foundin multiple myeloma, where the c-myc gene was found to have asuper-enhancer region that was bound by BRD4, giving rise to high levelsof c-myc transcription (see Loven, J., et al., “Selective Inhibition ofTumor Oncogenes by Disruption of Super-Enhancers” Cell 2013; 153,320-334, which is incorporated herein by reference in its entirety).

The MPLW515L mutation is found in a small percentage of patients withPhiladelphia-chromosome negative myeloproliferative neoplasms (MPN).This mutation causes constitutive activation of JAK2, leading toenhanced signaling through its downstream targets and subsequentincreased levels of cell proliferation. As there is much overlap betweenthe JAK/STAT signaling pathway and the inflammatory pathways regulatedtranscriptionally by the BET proteins, combination of BET and JAKinhibitors in MPN may form the bases for a new therapeutic treatmentprotocol.

There is a need for new therapies to improve patient outcome. Thisapplication is directed to this need and others.

SUMMARY

The present application provides, inter alia, methods of treating amyeloproliferative neoplasm in a patient in need thereof, comprisingadministering to said patient2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, and ruxolitinib, or apharmaceutically acceptable salt thereof.

The present application also provides methods of treating amyeloproliferative neoplasm in a patient in need thereof, comprisingadministering to said patient2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, and a dose of from about5 mg/day to about 60 mg/day on a free base basis of ruxolitinib, or apharmaceutically acceptable salt thereof.

The present application further provides the use of the compounds at thedoses recited herein for use in the manufacture of medicaments for usein treating a myeloproliferative neoplasm.

The present application also provides the compounds at the doses recitedherein for use in treating a myeloproliferative neoplasm.

The details of one or more embodiments are set forth in the descriptionbelow. Other features, objects, and advantages will be apparent from thedescription and from the claims.

DESCRIPTION OF THE FIGURES

FIG. 1A is a graph depicting efficacy of Compound 1 in the HEL xenograftmodel, tumor volume (mm³) vs. days post inoculation.

FIG. 1B is a graph depicting tolerability of Compound 1 in the HELxenograft model, tumor corrected body weight change (%) vs. days postinoculation.

FIG. 2A is a graph depicting efficacy of the combination of Compound 1and ruxolitinib in the SET-2 model, tumor volume (mm³) vs. days postinoculation.

FIG. 2B is a graph depicting tolerability of the combination of Compound1 and ruxolitinib in the SET-2 model, tumor corrected body weight change(%) vs. days post inoculation.

FIG. 3A is a graph depicting efficacy of the combination of Compound 1and ruxolitinib in the SET-2 model, tumor volume (mm³) vs. days postinoculation.

FIG. 3B is a graph depicting tolerability of the combination of Compound1 and ruxolitinib in the SET-2 model, tumor corrected body weight change(%) vs. days post inoculation.

FIG. 4A is a graph depicting spleen weight and white blood cell countsas measures of efficacy of Compound 1 and ruxolitinib in the MPLW515Lmouse model of MPN.

FIG. 4B is a graph depicting spleen weight and white blood cell countsas measures of efficacy of Compound 1 and ruxolitinib in the MPLW515Lmouse model of MPN.

FIG. 5 is a table (Table 3) depicting a summary of Compound 1pharmacokinetic parameters for Compound 1 as monotherapy (part 1 and 2)at steady-state (cycle 1 day 8).

FIG. 6 is a graph depicting inhibition of cMyc protein expression in thepharmacodynamics spiked cell assay versus plasma levels of Compound 1.

FIG. 7 is a graph depicting Compound 1 steady-state plasmaconcentrations (mean se) in participants following once daily dosing ofCompound 1 as monotherapy.

FIG. 8 is a graph depicting probability of response (hyperglycemia)versus Compound 1 steady-state AUC following once daily dosing ofCompound 1 as monotherapy.

DETAILED DESCRIPTION

For the terms “e.g.” and “such as,” and grammatical equivalents thereof,the phrase “and without limitation” is understood to follow unlessexplicitly stated otherwise.

As used herein, the singular forms “a,” “an,” and “the” include pluralreferents unless the context clearly dictates otherwise.

As used herein, the term “about” means “approximately” (e.g., plus orminus approximately 10% of the indicated value).

As used herein, the phrase “solid form” refers to a compound providedherein in either an amorphous state or a crystalline state (“crystallineform” or “crystalline solid” or “crystalline solid form”), whereby acompound provided herein in a crystalline state may optionally includesolvent or water within the crystalline lattice, for example, to form asolvated or hydrated crystalline form. The term “hydrated,” as usedherein, is meant to refer to a crystalline form that includes watermolecules in the crystalline lattice. Example “hydrated” crystallineforms include hemihydrates, monohydrates, dihydrates, and the like.Other hydrated forms such as channel hydrates and the like are alsoincluded within the meaning of the term.

The present invention relates to the use of a JAK1/JAK2 inhibitor,ruxolitinib, in combination with a BET protein inhibitor for treatmentof a myeloproliferative neoplasm. Ruxolitinib,(3R)-3-cyclopentyl-3-[4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]propanenitrile,is an inhibitor of JAK1 and JAK2. The IC₅₀ of ruxolitinib was measuredby Assay A infra at 1 mM ATP and found to be less than 10 nM at JAK1 andJAK2. Ruxolitinib can be made by the procedure described in U.S. Pat.No. 7,598,257 (Example 67), filed Dec. 12, 2006, which is incorporatedherein by reference in its entirety. Ruxolitinib phosphate can beprepared as described in US 2008/0312259, which is incorporated hereinby reference in its entirety.

It has been demonstrated that BET inhibition inhibits inflammatorysignaling in myeloproliferative neoplasms (see Kleppe M, Koche R, Zou L,et al., “Dual targeting of oncogenic activation and inflammatorysignaling increases therapeutic efficacy in myeloproliferativeneoplasms”. Cancer Cell 2018; 33:29-43.e7, which is incorporated hereinby reference in its entirety). In the referenced study, the BETinhibitor in combination with JAK inhibition reduced pathologic cytokineproduction and overall disease burden. Specifically, in an in vivo modelof myeloproliferative neoplasm, combination treatment with ruxolitiniband JQ1, a JAK1/2 and a BET inhibitor respectively, resulted indecreased inflammation, reduced disease burden. The treatment alsoeliminated fibrosis in myelofibrosis mice, a response not previouslyobserved with single-agent JAK inhibitor therapy. Mechanistically it isproposed that the synergistic effects of JAK and BET inhibitors aremediated by blocking 2 master regulators of pathologic inflammatorysignaling, JAK/STAT and NF-kB, respectively. Taken together, these datawarrant evaluation of BET inhibitors in malignancies characterized byunderlying inflammation, such as myeloproliferative neoplasms, includingprimary myelofibrosis.

The present application provides a method of treating amyeloproliferative neoplasm in a patient in need thereof, comprisingadministering to said patient2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one(Compound 1 below) is an inhibitor of BET proteins such as BRD2, BRD3,BRD4, and BRD-t. Compound 1 can be prepared as described in U.S. Pat.No. 9,540,368 or 10,189,832, each of which is incorporated herein byreference in its entirety.

The present application provides a method of treating amyeloproliferative neoplasm in a patient in need thereof, comprisingadministering to said patient2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, and ruxolitinib, or apharmaceutically acceptable salt thereof. Administering a combination ofCompound 1 and ruxolitinib can provide an enhanced efficacy. Asdemonstrated by the examples provided herein, combination of Compound 1and ruxolitinib showed enhanced efficacy in in vivo models andsignificantly (p<0.05) more efficacious than either ruxolitinib alone orCompound 1 alone. That is, the combination of ruxolitinib and Compound 1provides efficacy better than the sum of the parts.

The present application provides a method of treating amyeloproliferative neoplasm in a patient in need thereof, comprisingadministering to said patient2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, and ruxolitinib, or apharmaceutically acceptable salt thereof.

The present application further provides a method of treating amyeloproliferative neoplasm in a patient in need thereof, comprisingadministering to said patient2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, and a therapeuticallyeffective amount of ruxolitinib, or a pharmaceutically acceptable saltthereof.

The present application further provides a method of treating amyeloproliferative neoplasm in a patient in need thereof, comprisingadministering to said patient a dose from about 2 mg/day to about 20mg/day on a free base basis of2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, and a therapeuticallyeffective amount of ruxolitinib, or a pharmaceutically acceptable saltthereof.

In some embodiments, the dose of2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, is about 2 mg/day toabout 18 mg/day on a free base basis.

In some embodiments, the dose of2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, is about 2 mg/day toabout 12 mg/day on a free base basis.

In some embodiments, the dose of2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, is about 4 mg/day toabout 8 mg/day on a free base basis.

For example, the dose of2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, is about 2, about 4,about 6, about 10, about 12, about 14, about 16, about 18, or about 20mg/day on a free base basis. In some embodiments, the dose of2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, is about 2, about 4,about 6, about 8, about 10, about 12, about 14, about 16, about 18, orabout 20 mg/day on a free base basis. In some embodiments, the dose of2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, is about 2, about 4,about 6, about 8, about 10, or about 12 mg per day on a free base basis.In some embodiments, the dose of2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, is about 4, about 6, orabout 8 mg per day on a free base basis. In some embodiments, the doseof2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, is about 8, about 10,about 12, about 14, or about 16 mg per day on a free base basis. In someembodiments, the dose of2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, is about 10, about 12,about 14, about 16 mg, or about 18 mg per day on a free base basis. Insome embodiments, the dose of2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, is about 2 mg per day ona free base basis. In some embodiments, the dose of2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, is about 4 mg per day ona free base basis. In some embodiments, the dose of2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, is about 6 mg per day ona free base basis. In some embodiments, the dose of2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, is about 8 mg per day ona free base basis. In some embodiments, the dose of2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, is about 10 mg per day ona free base basis. In some embodiments, the dose of2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, is about 12 mg per day ona free base basis.

In some embodiments,2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, and ruxolitinib, or apharmaceutically acceptable salt thereof are administered once daily(QD). In some embodiments,2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, and ruxolitinib, or apharmaceutically acceptable salt thereof are administered twice daily(BID).

In some embodiments, the dose of ruxolitinib, or a pharmaceuticallyacceptable salt thereof, is about 5 mg/day to about 60 mg/day on a freebase basis. For example, the dose of ruxolitinib, or a pharmaceuticallyacceptable salt thereof, is about 5, about 10, about 15, about 20, about25, about 30, about 35, about 40, about 45, about 50, about 55, or 60mg/day on a free base basis.

In some embodiments, the dose of ruxolitinib, or a pharmaceuticallyacceptable salt thereof, is about 2.5 mg BID to about 30 mg BID on afree base basis. For example, the dose of ruxolitinib, or apharmaceutically acceptable salt thereof, is about 2.5, about 5, about7.5, about 10, about 12.5, about 15, about 17.5, about 20, about 25, orabout 30 mg BID on a free base basis.

In some embodiments, the pharmaceutically acceptable salt of ruxolitinibis ruxolitinib phosphate.

In some embodiments,2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, is selected from thephosphoric acid salt, dihydrochloric acid salt, hydrochloric acid salt,maleic acid salt, adipic acid salt, hydrobromic acid salt,(R)-(−)-mandelic acid salt, salicylic acid salt, benzoic acid salt,benzenesulfonic acid salt, L-pyroglutamic acid salt, methanesulfonicacid salt, (1S)-(+)-10-camphorsulfonic acid salt, fumaric acid salt,sulfuric acid salt, L-tartaric acid salt, and D-tartaric acid salt of2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one.In some embodiments,2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-oneis a free base.

In some embodiments, the2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-oneincludes one or more crystalline solid forms of an inhibitor of a BETprotein. A more detailed discussion of Compound 1's crystalline formscan be found in U.S. Pat. No. 10,626,114, which is incorporated hereinby reference in its entirety, and briefly described herein. Typically,different crystalline forms of the same substance have different bulkproperties relating to, for example, hygroscopicity, solubility,stability, and the like.

The crystalline solid forms of Compound 1 can include solvent such aswater (e.g., a hydrated form) or be substantially free of water andsolvent (e.g., forming an anhydrate). In some embodiments, thecrystalline solid form is an anhydrate. In further embodiments, thecrystalline solid form is hydrated. Compound 1 can be obtained in asolid crystalline form referred to as Form I, Form II, Form III, FormIV, Form V, Form Va, Form VI, Form VII, Form VIII, Form IX, Form X, FormXI, Form XII, Form XIII, Form XIV, and Form X, which are described inU.S. Pat. No. 10,626,114, which is incorporated herein by reference inits entirety.

Experimental data show that Form I is an anhydrate. Form I ischaracterized by its XRPD pattern and other solid state characteristics.In some embodiments, Form I has one or more characteristic XRPD peaks,in terms of 2-theta, selected from about 8.7, about 9.8, about 12.7,about 21.4, and about 23.3 degrees.

Experimental data show that Form II is an anhydrate. Form II ischaracterized by its XRPD pattern and other solid state characteristics.In some embodiments, Form II has one or more characteristic XRPD peaks,in terms of 2-theta, selected from about 6.7, about 9.5, about 10.5,about 14.8, about 16.2, about 17.0, about 18.8, and about 19.3 degrees.

In some embodiments, Form III has one or more characteristic XRPD peaks,in terms of 2-theta, selected from about 7.8, about 12.4, about 13.1,about 15.2, about 15.5, about 16.9, about 17.5, and about 20.3 degrees.

In some embodiments, Form IV has one or more characteristic XRPD peaks,in terms of 2-theta, selected from about 11.2, about 16.3, about 18.7,and about 22.1 degrees.

In some embodiments, Form V has one or more characteristic XRPD peaks,in terms of 2-theta, selected from about 8.2, about 8.5, about 14.1,about 16.3, about 17.1, about 18.9, about 19.8, about 21.8, and about22.7 degrees.

In some embodiments, Form Va has one or more characteristic XRPD peaks,in terms of 2-theta, selected from about 8.7, about 16.5, about 17.3,about 19.9, and about 21.6 degrees.

In some embodiments, Form VI has one or more characteristic XRPD peaks,in terms of 2-theta, selected from about 8.5, about 9.6, about 11.4,about 12.1, about 13.5, about 14.5, about 15.2, about 17.1, about 17.7,about 18.1, about 19.2, and about 20.7 degrees.

In some embodiments, Form VII has one or more characteristic XRPD peaks,in terms of 2-theta, selected from about 9.9, about 12.2, about 14.8,about 15.7, about 17.0, about 17.5, and about 18.8 degrees.

In some embodiments, Form VIII has one or more characteristic XRPDpeaks, in terms of 2-theta, selected from about 8.1, about 8.5, about16.2, about 16.6, about 17.0, about 17.5, about 18.0, about 18.9, about19.6, and about 20.1 degrees.

In some embodiments, Form IX has one or more characteristic XRPD peaks,in terms of 2-theta, selected from about 8.6, about 9.1, about 11.4,about 13.4, about 15.2, about 18.2, about 22.1, about 22.8, and about23.9 degrees.

In some embodiments, Form X has one or more characteristic XRPD peaks,in terms of 2-theta, selected from about 14.9, about 15.3, about 15.8,about 17.0, about 17.7, about 18.3, and about 19.7 degrees.

In some embodiments, Form XI has one or more characteristic XRPD peaks,in terms of 2-theta, selected from about 8.9, about 12.8, about 18.0about 21.5, about 22.6, and about 23.3 degrees.

In some embodiments, Form XII has one or more characteristic XRPD peaks,in terms of 2-theta, selected from about 5.6, about 11.7, about 13.8,about 14.5, about 16.9, about 17.7, about 18.7, about 23.5, about 24.6,about 34.3, about 44.2, and 44.6 degrees.

In some embodiments, Form XIII has one or more characteristic XRPDpeaks, in terms of 2-theta, selected from about 5.7, about 8.6, about9.8, about 11.8, about 12.6, about 13.4, about 14.1, about 14.8, about16.6, and about 19.1 degrees.

In some embodiments, Form XIV has one or more characteristic XRPD peaks,in terms of 2-theta, selected from about 4.0, about 11.2, about 11.9,about 14.1, about 14.8, and about 15.9 degrees.

In some embodiments, Form XV has one or more characteristic XRPD peaks,in terms of 2-theta, selected from about 7.4, about 9.6, about 12.4,about 13.4, about 15.5, about 16.9, about 17.7, about 19.0, about 19.5,about 20.6, and about 22.5 degrees.

The present application provides a method of treating amyeloproliferative neoplasm in a patient in need thereof, comprisingadministering to said patient2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, and a therapeuticallyeffective amount of ruxolitinib, or a pharmaceutically acceptable saltthereof. In some embodiments, the myeloproliferative neoplasm isselected from polycythemia vera (PV), essential thrombocythemia (ET),primary myelofibrosis, chronic myelogenous leukemia (CML), chronicmyelomonocytic leukemia (CMML), hypereosinophilic syndrome (HES),systemic mast cell disease (SMCD), chronic neutrophilic leukemia (CNL),and chronic eosinophilic leukemia.

In some embodiments, the myeloproliferative neoplasm is polycythemiavera (PV).

In some embodiments, the myeloproliferative neoplasm is essentialthrombocythemia (ET).

In some embodiments, the myeloproliferative neoplasm is primarymyelofibrosis.

In some embodiments, the myeloproliferative neoplasm is myelofibrosis.

In some embodiments, ruxolitinib, or pharmaceutically acceptable saltthereof, and2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, are administered orally.In some embodiments, the ruxolitinib, or pharmaceutically acceptablesalt thereof, is administered orally. In some embodiments,2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, is administered orally.

In some embodiments, ruxolitinib, or pharmaceutically acceptable saltthereof, and2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, are administeredsimultaneously or sequentially.

The preceding embodiments are intended to be combined in any suitablecombination as if the embodiments are multiply dependent claims (e.g.,the embodiments related to the individual doses for ruxolitinib, theembodiments related to the individual doses for the BET proteininhibitor (Compound 1), the embodiments related to the salt forms, theembodiments related to the individual types of myeloproliferativeneoplasms, and the embodiments related to oral administration can becombined in any combination). The combinations are not separately listedherein merely for the sake of brevity.

All compounds, and pharmaceutically acceptable salts thereof, can befound together with other substances such as water and solvents (e.g.,hydrates and solvates) or can be isolated. When in the solid state, thecompounds described herein and salts thereof may occur in various formsand may, e.g., take the form of solvates, including hydrates. Thecompounds may be in any solid state form, such as a polymorph orsolvate, so unless clearly indicated otherwise, reference in thespecification to compounds and salts thereof should be understood asencompassing any solid state form of the compound.

The phrase “pharmaceutically acceptable” is employed herein to refer tothose compounds, materials, compositions and/or dosage forms which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of human beings and animals without excessive toxicity,irritation, allergic response, or other problem or complication,commensurate with a reasonable benefit/risk ratio.

The present invention also includes pharmaceutically acceptable salts ofthe compounds described herein. The term “pharmaceutically acceptablesalts” refers to derivatives of the disclosed compounds wherein theparent compound is modified by converting an existing acid or basemoiety to its salt form. Examples of pharmaceutically acceptable saltsinclude, but are not limited to, mineral or organic acid salts of basicresidues such as amines; alkali or organic salts of acidic residues suchas carboxylic acids; and the like. The pharmaceutically acceptable saltsof the present invention include the non-toxic salts of the parentcompound formed, e.g., from non-toxic inorganic or organic acids. Thepharmaceutically acceptable salts of the present invention can besynthesized from the parent compound which contains a basic or acidicmoiety by conventional chemical methods. Generally, such salts can beprepared by reacting the free acid or base forms of these compounds witha stoichiometric amount of the appropriate base or acid in water or inan organic solvent, or in a mixture of the two; generally, non-aqueousmedia like ether, ethyl acetate, alcohols (e.g., methanol, ethanol,iso-propanol, or butanol) or acetonitrile (MeCN) are preferred. Lists ofsuitable salts are found in Remington's Pharmaceutical Sciences, 17^(th)Ed., (Mack Publishing Company, Easton, 1985), p. 1418, Berge et al., J.Pharm. Sci., 1977, 66(1), 1-19, and in Stahl et al., Handbook ofPharmaceutical Salts: Properties, Selection, and Use, (Wiley, 2002). Insome embodiments, the compounds described herein include the N-oxideforms.

The dosages described herein are on a free base basis. The phrase “on afree base basis” indicates that the amount of the compound (e.g.,ruxolitinib or salt thereof) in the dosage form is measured based on themolecular weight of the compound free base only, even when the actualactive ingredient is a salt of the compound having a different molecularweight than the free base. For example, the conversion factor forruxolitinib phosphate salt to free base is 0.7575.

The terms “individual” or “patient,” used interchangeably, refer to anyanimal, including mammals, preferably mice, rats, other rodents,rabbits, dogs, cats, swine, cattle, sheep, horses, or primates, and mostpreferably humans.

The phrase “therapeutically effective amount” refers to the amount ofactive compound or pharmaceutical agent that elicits the biological ormedicinal response in a tissue, system, animal, individual or human thatis being sought by a researcher, veterinarian, medical doctor or otherclinician.

The term “treating” or “treatment” refers to one or more of (1)inhibiting the disease; e.g., inhibiting a disease, condition ordisorder in an individual who is experiencing or displaying thepathology or symptomatology of the disease, condition or disorder (i.e.,arresting further development of the pathology and/or symptomatology);and (2) ameliorating the disease; e.g., ameliorating a disease,condition or disorder in an individual who is experiencing or displayingthe pathology or symptomatology of the disease, condition or disorder(i.e., reversing the pathology and/or symptomatology) such as decreasingthe severity of disease. In one embodiment, treating or treatmentincludes preventing or reducing the risk of developing the disease;e.g., preventing or reducing the risk of developing a disease, conditionor disorder in an individual who may be predisposed to the disease,condition or disorder but does not yet experience or display thepathology or symptomatology of the disease.

The term “BID” means two times a day.

The term “QD” means once a day.

Additional Combinations

Compound 1 can be used in additional combination treatments whereCompound 1 is administered in conjunction with other treatments such asthe administration of one or more additional therapeutic agents. Theadditional therapeutic agents are typically those which are normallyused to treat the particular condition to be treated. The additionaltherapeutic agents can include, e.g., chemotherapeutics,anti-inflammatory agents, steroids, immunosuppressants, as well asBcr-Abl, Flt-3, RAF, FAK, and JAK kinase inhibitors for treatment of BETprotein-associated diseases, disorders or conditions. The one or moreadditional pharmaceutical agents can be administered to a patientsimultaneously or sequentially.

In some embodiments, Compound 1 can be used in combination with atherapeutic agent that targets an epigenetic regulator. Examples ofepigenetic regulators include the histone lysine methyltransferases,histone arginine methyl transferases, histone demethylases, histonedeacetylases, histone acetylases, and DNA methyltransferases. Histonedeacetylase inhibitors include, e.g., vorinostat.

For treating cancer and other proliferative diseases, Compound 1 can beused in combination with chemotherapeutic agents, or otheranti-proliferative agents. Compound 1 can also be used in combinationwith medical therapy such as surgery or radiotherapy, e.g.,gamma-radiation, neutron beam radiotherapy, electron beam radiotherapy,proton therapy, brachytherapy, and systemic radioactive isotopes.Examples of suitable chemotherapeutic agents include any of: abarelix,aldesleukin, alemtuzumab, alitretinoin, allopurinol, altretamine,anastrozole, arsenic trioxide, asparaginase, azacitidine, bevacizumab,bexarotene, bleomycin, bortezombi, bortezomib, busulfan intravenous,busulfan oral, calusterone, capecitabine, carboplatin, carmustine,cetuximab, chlorambucil, cisplatin, cladribine, clofarabine,cyclophosphamide, cytarabine, dacarbazine, dactinomycin, dalteparinsodium, dasatinib, daunorubicin, decitabine, denileukin, denileukindiftitox, dexrazoxane, docetaxel, doxorubicin, dromostanolonepropionate, eculizumab, epirubicin, erlotinib, estramustine, etoposidephosphate, etoposide, exemestane, fentanyl citrate, filgrastim,floxuridine, fludarabine, fluorouracil, fulvestrant, gefitinib,gemcitabine, gemtuzumab ozogamicin, goserelin acetate, histrelinacetate, ibritumomab tiuxetan, idarubicin, ifosfamide, imatinibmesylate, interferon alfa 2a, irinotecan, lapatinib ditosylate,lenalidomide, letrozole, leucovorin, leuprolide acetate, levamisole,lomustine, meclorethamine, megestrol acetate, melphalan, mercaptopurine,methotrexate, methoxsalen, mitomycin C, mitotane, mitoxantrone,nandrolone phenpropionate, nelarabine, nofetumomab, oxaliplatin,paclitaxel, pamidronate, panitumumab, pegaspargase, pegfilgrastim,pemetrexed disodium, pentostatin, pipobroman, plicamycin, procarbazine,quinacrine, rasburicase, rituximab, ruxolitinib, sorafenib,streptozocin, sunitinib, sunitinib maleate, tamoxifen, temozolomide,teniposide, testolactone, thalidomide, thioguanine, thiotepa, topotecan,toremifene, tositumomab, trastuzumab, tretinoin, uracil mustard,valrubicin, vinblastine, vincristine, vinorelbine, vorinostat, andzoledronate.

For treating cancer and other proliferative diseases, Compound 1 can beused in combination with ruxolitinib.

Compound 1 can be used in combination with one or more immune checkpointinhibitors. Exemplary immune checkpoint inhibitors include inhibitorsagainst immune checkpoint molecules such as CD27, CD28, CD40, CD122,CD96, CD73, CD47, OX40, GITR, CSF1R, JAK, PI3K delta, PI3K gamma, TAM,arginase, CD137 (also known as 4-1BB), ICOS, A2AR, B7-H3, B7-H4, BTLA,CTLA-4, LAG3, TIM3, VISTA, PD-1, PD-L1 and PD-L2. In some embodiments,the immune checkpoint molecule is a stimulatory checkpoint moleculeselected from CD27, CD28, CD40, ICOS, OX40, GITR and CD137. In someembodiments, the immune checkpoint molecule is an inhibitory checkpointmolecule selected from A2AR, B7-H3, B7-H4, BTLA, CTLA-4, IDO, KIR, LAG3,PD-1, TIM3, and VISTA. In some embodiments, the compounds providedherein can be used in combination with one or more agents selected fromKIR inhibitors, TIGIT inhibitors, LAIR1 inhibitors, CD160 inhibitors,2B4 inhibitors and TGFR beta inhibitors.

In some embodiments, the inhibitor of an immune checkpoint molecule isanti-PD1 antibody, anti-PD-L1 antibody, or anti-CTLA-4 antibody.

In some embodiments, the inhibitor of an immune checkpoint molecule isan inhibitor of PD-1, e.g., an anti-PD-1 monoclonal antibody. In someembodiments, the anti-PD-1 monoclonal antibody is nivolumab,pembrolizumab (also known as MK-3475), pidilizumab, SHR-1210, PDR001, orAMP-224. In some embodiments, the anti-PD-1 monoclonal antibody isnivolumab or pembrolizumab. In some embodiments, the anti-PD1 antibodyis pembrolizumab.

In some embodiments, the inhibitor of an immune checkpoint molecule isan inhibitor of PD-L1, e.g., an anti-PD-L1 monoclonal antibody. In someembodiments, the anti-PD-L1 monoclonal antibody is BMS-935559, MEDI4736,MPDL3280A (also known as RG7446), or MSB0010718C. In some embodiments,the anti-PD-L1 monoclonal antibody is MPDL3280A or MEDI4736.

In some embodiments, the inhibitor of an immune checkpoint molecule isan inhibitor of CTLA-4, e.g., an anti-CTLA-4 antibody. In someembodiments, the anti-CTLA-4 antibody is ipilimumab.

In some embodiments, the inhibitor of an immune checkpoint molecule isan inhibitor of LAG3, e.g., an anti-LAG3 antibody. In some embodiments,the anti-LAG3 antibody is BMS-986016 or LAG525.

In some embodiments, the inhibitor of an immune checkpoint molecule isan inhibitor of GITR, e.g., an anti-GITR antibody. In some embodiments,the anti-GITR antibody is TRX518 or MK-4166.

In some embodiments, the inhibitor of an immune checkpoint molecule isan inhibitor of OX40, e.g., an anti-OX40 antibody or OX40L fusionprotein. In some embodiments, the anti-OX40 antibody is MEDI0562. Insome embodiments, the OX40L fusion protein is MED16383.

Compound 1 can be used in combination with one or more agents for thetreatment of diseases such as cancer. In some embodiments, the agent isan alkylating agent, a proteasome inhibitor, a corticosteroid, or animmunomodulatory agent. Examples of an alkylating agent includecyclophosphamide (CY), melphalan (MEL), and bendamustine. In someembodiments, the proteasome inhibitor is carfilzomib. In someembodiments, the corticosteroid is dexamethasone (DEX). In someembodiments, the immunomodulatory agent is lenalidomide (LEN) orpomalidomide (POM).

For treating autoimmune or inflammatory conditions, Compound 1 can beadministered in combination with a corticosteroid such as triamcinolone,dexamethasone, fluocinolone, cortisone, prednisolone, or flumetholone.

For treating autoimmune or inflammatory conditions, Compound 1 can beadministered in combination with an immune suppressant such asfluocinolone acetonide (Retisert®), rimexolone (AL-2178, Vexol, Alcon),or cyclosporine (Restasis®).

For treating autoimmune or inflammatory conditions, Compound 1 can beadministered in combination with one or more additional agents selectedfrom Dehydrex™ (Holles Labs), Civamide (Opko), sodium hyaluronate(Vismed, Lantibio/TRB Chemedia), cyclosporine (ST-603, SirionTherapeutics), ARG101(T) (testosterone, Argentis), AGR1012(P)(Argentis), ecabet sodium (Senju-Ista), gefamate (Santen),15-(s)-hydroxyeicosatetraenoic acid (15(S)-HETE), cevilemine,doxycycline (ALTY-0501, Alacrity), minocycline, iDestrin™ (NP50301,Nascent Pharmaceuticals), cyclosporine A (Nova22007, Novagali),oxytetracycline (Duramycin, MOLI1901, Lantibio), CF101 (2S,3S, 4R,5R)-3,4-dihydroxy-5-[6-[(3-iodophenyl)methylamino]purin-9-yl]-N-methyl-oxolane-2-carbamyl,Can-Fite Biopharma), voclosporin (LX212 or LX214, Lux Biosciences),ARG103 (Agentis), RX-10045 (synthetic resolvin analog, Resolvyx), DYN15(Dyanmis Therapeutics), rivoglitazone (DE011, Daiichi Sanko), TB4(RegeneRx), OPH-01 (Ophtalmis Monaco), PCS101 (Pericor Science), REV1-31(Evolutec), Lacritin (Senju), rebamipide (Otsuka-Novartis), OT-551(Othera), PAI-2 (University of Pennsylvania and Temple University),pilocarpine, tacrolimus, pimecrolimus (AMS981, Novartis), loteprednoletabonate, rituximab, diquafosol tetrasodium (INS365, Inspire), KLS-0611(Kissei Pharmaceuticals), dehydroepiandrosterone, anakinra, efalizumab,mycophenolate sodium, etanercept (Embrel®), hydroxychloroquine, NGX267(TorreyPines Therapeutics), or thalidomide.

In some embodiments, Compound 1 can be administered in combination withone or more agents selected from an antibiotic, antiviral, antifungal,anesthetic, anti-inflammatory agents including steroidal andnon-steroidal anti-inflammatories, and anti-allergic agents. Examples ofsuitable medicaments include aminoglycosides such as amikacin,gentamycin, tobramycin, streptomycin, netilmycin, and kanamycin;fluoroquinolones such as ciprofloxacin, norfloxacin, ofloxacin,trovafloxacin, lomefloxacin, levofloxacin, and enoxacin; naphthyridine;sulfonamides; polymyxin; chloramphenicol; neomycin; paramomycin;colistimethate; bacitracin; vancomycin; tetracyclines; rifampin and itsderivatives (“rifampins”); cycloserine; beta-lactams; cephalosporins;amphotericins; fluconazole; flucytosine; natamycin; miconazole;ketoconazole; corticosteroids; diclofenac; flurbiprofen; ketorolac;suprofen; cromolyn; lodoxamide; levocabastin; naphazoline; antazoline;pheniramine; or azalide antibiotic.

Other examples of agents, one or more of which a provided compound mayalso be combined with include: a treatment for Alzheimer's Disease suchas donepezil and rivastigmine; a treatment for Parkinson's Disease suchas L-DOPA/carbidopa, entacapone, ropinirole, pramipexole, bromocriptine,pergolide, trihexyphenidyl, and amantadine; an agent for treatingmultiple sclerosis (MS) such as beta interferon (e.g., Avonex® andRebif®), glatiramer acetate, and mitoxantrone; a treatment for asthmasuch as albuterol and montelukast; an agent for treating schizophreniasuch as zyprexa, risperdal, seroquel, and haloperidol; ananti-inflammatory agent such as a corticosteroid, such as dexamethasoneor prednisone, a TNF blocker, IL-1 RA, azathioprine, cyclophosphamide,and sulfasalazine; an immunomodulatory agent, includingimmunosuppressive agents, such as cyclosporin, tacrolimus, rapamycin,mycophenolate mofetil, an interferon, a corticosteroid,cyclophosphamide, azathioprine, and sulfasalazine; a neurotrophic factorsuch as an acetylcholinesterase inhibitor, an MAO inhibitor, aninterferon, an anti-convulsant, an ion channel blocker, riluzole, or ananti-Parkinson's agent; an agent for treating cardiovascular diseasesuch as a beta-blocker, an ACE inhibitor, a diuretic, a nitrate, acalcium channel blocker, or a statin; an agent for treating liverdisease such as a corticosteroid, cholestyramine, an interferon, and ananti-viral agent; an agent for treating blood disorders such as acorticosteroid, an anti-leukemic agent, or a growth factor; or an agentfor treating immunodeficiency disorders such as gamma globulin.

In some embodiments, Compound 1 is administered in combination with aJAK kinase inhibitor (e.g., ruxolitinib, tofacitinib, baricitinib,CYT387, GLPG0634, lestaurtinib, pacritinib, TG101348, or aJAK1-selective inhibitor), a Pim kinase inhibitor (including inhibitorsof one or more of PIM1, PIM2, and PIM3), a PI3 kinase inhibitorincluding PI3K-delta selective and broad spectrum PI3K inhibitors, anMEK inhibitor, a cyclin dependent kinase inhibitor, a b-RAF inhibitor,an mTOR inhibitor, a proteasome inhibitor (e.g., bortezomib,carfilzomib), an HDAC-inhibitor (e.g., panobinostat, vorinostat), a DNAmethyl transferase inhibitor, dexamethasone, melphalan, or animmunomodulator (e.g., lenolidomide, pomalidomide).

When more than one pharmaceutical agent is administered to a patient,they can be administered simultaneously, sequentially, or in combination(e.g., for more than two agents).

Compositions

The compounds can be administered in the form of pharmaceuticalcompositions. These compositions can be prepared in a manner well knownin the pharmaceutical art, and can be administered by a variety ofroutes, depending upon whether local or systemic treatment is indicatedand upon the area to be treated. Administration may be topical(including transdermal, epidermal, ophthalmic and to mucous membranesincluding intranasal, vaginal and rectal delivery), pulmonary (e.g., byinhalation or insufflation of powders or aerosols, including bynebulizer; intratracheal or intranasal), oral or parenteral. Parenteraladministration includes intravenous, intraarterial, subcutaneous,intraperitoneal intramuscular or injection or infusion; or intracranial,e.g., intrathecal or intraventricular, administration. Parenteraladministration can be in the form of a single bolus dose, or may be,e.g., by a continuous perfusion pump. Pharmaceutical compositions andformulations for topical administration may include transdermal patches,ointments, lotions, creams, gels, drops, suppositories, sprays, liquidsand powders. Conventional pharmaceutical carriers, aqueous, powder oroily bases, thickeners and the like may be necessary or desirable.

The pharmaceutical compositions can contain, as the active ingredient,the compounds, or a pharmaceutically acceptable salt thereof, incombination with one or more pharmaceutically acceptable carriers(excipients). In some embodiments, the composition is suitable fortopical administration. In making the compositions, the activeingredient is typically mixed with an excipient, diluted by an excipientor enclosed within such a carrier in the form of, e.g., a capsule,sachet, paper, or other container. When the excipient serves as adiluent, it can be a solid, semi-solid, or liquid material, which actsas a vehicle, carrier or medium for the active ingredient. Thus, thecompositions can be in the form of tablets, pills, powders, lozenges,sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups,aerosols (as a solid or in a liquid medium), ointments containing, e.g.,up to 10% by weight of the active compound, soft and hard gelatincapsules, suppositories, sterile injectable solutions and sterilepackaged powders.

In preparing a formulation, the active compound can be milled to providethe appropriate particle size prior to combining with the otheringredients. If the active compound is substantially insoluble, it canbe milled to a particle size of less than 200 mesh. If the activecompound is substantially water soluble, the particle size can beadjusted by milling to provide a substantially uniform distribution inthe formulation, e.g., about 40 mesh.

The compounds may be milled using known milling procedures such as wetmilling to obtain a particle size appropriate for tablet formation andfor other formulation types. Finely divided (nanoparticulate)preparations of the compounds of the invention can be prepared byprocesses known in the art see, e.g., WO 2002/000196.

Some examples of suitable excipients include lactose, dextrose, sucrose,sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates,tragacanth, gelatin, calcium silicate, microcrystalline cellulose,polyvinylpyrrolidone, cellulose, water, syrup and methyl cellulose. Theformulations can additionally include: lubricating agents such as talc,magnesium stearate and mineral oil; wetting agents; emulsifying andsuspending agents; preserving agents such as methyl- andpropylhydroxy-benzoates; and sweetening agents and flavoring agents. Thecompositions of the invention can be formulated so as to provide quick,sustained or delayed release of the active ingredient afteradministration to the patient by employing procedures known in the art.

In some embodiments, the pharmaceutical composition comprises silicifiedmicrocrystalline cellulose (SMCC) and at least one compound describedherein, or a pharmaceutically acceptable salt thereof. In someembodiments, the silicified microcrystalline cellulose comprises about98% microcrystalline cellulose and about 2% silicon dioxide w/w.

In some embodiments, the composition is a sustained release compositioncomprising at least one compound described herein, or a pharmaceuticallyacceptable salt thereof, and at least one pharmaceutically acceptablecarrier. In some embodiments, the composition comprises at least onecompound described herein, or a pharmaceutically acceptable saltthereof, and at least one component selected from microcrystallinecellulose, lactose monohydrate, hydroxypropyl methylcellulose andpolyethylene oxide. In some embodiments, the composition comprises atleast one compound described herein, or a pharmaceutically acceptablesalt thereof, and microcrystalline cellulose, lactose monohydrate andhydroxypropyl methylcellulose. In some embodiments, the compositioncomprises at least one compound described herein, or a pharmaceuticallyacceptable salt thereof, and microcrystalline cellulose, lactosemonohydrate and polyethylene oxide. In some embodiments, the compositionfurther comprises magnesium stearate or silicon dioxide. In someembodiments, the microcrystalline cellulose is Avicel PH102™. In someembodiments, the lactose monohydrate is Fast-flo 316™. In someembodiments, the hydroxypropyl methylcellulose is hydroxypropylmethylcellulose 2208 K4M (e.g., Methocel K4 M Premier™) and/orhydroxypropyl methylcellulose 2208 K100LV (e.g., Methocel KOOLV™) Insome embodiments, the polyethylene oxide is polyethylene oxide WSR 1105(e.g., Polyox WSR 1105™).

The components used to formulate the pharmaceutical compositions are ofhigh purity and are substantially free of potentially harmfulcontaminants (e.g., at least National Food grade, generally at leastanalytical grade, and more typically at least pharmaceutical grade).Particularly for human consumption, the composition is preferablymanufactured or formulated under Good Manufacturing Practice standardsas defined in the applicable regulations of the U.S. Food and DrugAdministration. For example, suitable formulations may be sterile and/orsubstantially isotonic and/or in full compliance with all GoodManufacturing Practice regulations of the U.S. Food and DrugAdministration.

The active compound may be effective over a wide dosage range and isgenerally administered in a therapeutically effective amount. It will beunderstood, however, that the amount of the compound actuallyadministered will usually be determined by a physician, according to therelevant circumstances, including the condition to be treated, thechosen route of administration, the actual compound administered, theage, weight and response of the individual patient, the severity of thepatient's symptoms and the like.

The therapeutic dosage of a compound of the present invention can varyaccording to, e.g., the particular use for which the treatment is made,the manner of administration of the compound, the health and conditionof the patient, and the judgment of the prescribing physician. Theproportion or concentration of a compound of the invention in apharmaceutical composition can vary depending upon a number of factorsincluding dosage, chemical characteristics (e.g., hydrophobicity), andthe route of administration.

For preparing solid compositions such as tablets, the principal activeingredient is mixed with a pharmaceutical excipient to form a solidpreformulation composition containing a homogeneous mixture of acompound of the present invention. When referring to thesepreformulation compositions as homogeneous, the active ingredient istypically dispersed evenly throughout the composition so that thecomposition can be readily subdivided into equally effective unit dosageforms such as tablets, pills and capsules. This solid preformulation isthen subdivided into unit dosage forms of the type described abovecontaining from, e.g., about 0.1 to about 1000 mg of the activeingredient of the present invention.

The tablets or pills of the present invention can be coated or otherwisecompounded to provide a dosage form affording the advantage of prolongedaction. For example, the tablet or pill can comprise an inner dosage andan outer dosage component, the latter being in the form of an envelopeover the former. The two components can be separated by an enteric layerwhich serves to resist disintegration in the stomach and permit theinner component to pass intact into the duodenum or to be delayed inrelease. A variety of materials can be used for such enteric layers orcoatings, such materials including a number of polymeric acids andmixtures of polymeric acids with such materials as shellac, cetylalcohol and cellulose acetate.

The liquid forms in which the compounds and compositions of the presentinvention can be incorporated for administration orally or by injectioninclude aqueous solutions, suitably flavored syrups, aqueous or oilsuspensions, and flavored emulsions with edible oils such as cottonseedoil, sesame oil, coconut oil, or peanut oil, as well as elixirs andsimilar pharmaceutical vehicles.

Compositions for inhalation or insufflation include solutions andsuspensions in pharmaceutically acceptable, aqueous or organic solvents,or mixtures thereof, and powders. The liquid or solid compositions maycontain suitable pharmaceutically acceptable excipients as describedsupra. In some embodiments, the compositions are administered by theoral or nasal respiratory route for local or systemic effect.Compositions can be nebulized by use of inert gases. Nebulized solutionsmay be breathed directly from the nebulizing device or the nebulizingdevice can be attached to a face mask, tent, or intermittent positivepressure breathing machine. Solution, suspension, or powder compositionscan be administered orally or nasally from devices which deliver theformulation in an appropriate manner.

Topical formulations can contain one or more conventional carriers. Insome embodiments, ointments can contain water and one or morehydrophobic carriers selected from, e.g., liquid paraffin,polyoxyethylene alkyl ether, propylene glycol, white Vaseline®(petroleum jelly) and the like. Carrier compositions of creams can bebased on water in combination with glycerol and one or more othercomponents, e.g., glycerinemonostearate, PEG-glycerinemonostearate andcetylstearyl alcohol. Gels can be formulated using isopropyl alcohol andwater, suitably in combination with other components such as, e.g.,glycerol, hydroxyethyl cellulose and the like.

The amount of compound or composition administered to a patient willvary depending upon what is being administered, the purpose of theadministration, such as prophylaxis or therapy, the state of thepatient, the manner of administration and the like. In therapeuticapplications, compositions can be administered to a patient alreadysuffering from a disease in an amount sufficient to cure or at leastpartially arrest the symptoms of the disease and its complications.Effective doses will depend on the disease condition being treated aswell as by the judgment of the attending clinician depending uponfactors such as the severity of the disease, the age, weight and generalcondition of the patient and the like.

The compositions administered to a patient can be in the form ofpharmaceutical compositions described above. These compositions can besterilized by conventional sterilization techniques, or may be sterilefiltered. Aqueous solutions can be packaged for use as is, orlyophilized, the lyophilized preparation being combined with a sterileaqueous carrier prior to administration. The pH of the compoundpreparations typically will be between 3 and 11, more preferably from 5to 9 and most preferably from 7 to 8. It will be understood that use ofcertain of the foregoing excipients, carriers or stabilizers will resultin the formation of pharmaceutical salts.

Kits

The present application also includes pharmaceutical kits useful, whichinclude one or more containers containing a pharmaceutical compositioncomprising a therapeutically effective amount of the compound, or any ofthe embodiments thereof. Such kits can further include one or more ofvarious conventional pharmaceutical kit components, such as, e.g.,containers with one or more pharmaceutically acceptable carriers,additional containers, etc., as will be readily apparent to thoseskilled in the art. Instructions, either as inserts or as labels,indicating quantities of the components to be administered, guidelinesfor administration, and/or guidelines for mixing the components, canalso be included in the kit.

The invention will be described in greater detail by way of specificexamples. The following examples are offered for illustrative purposes,and are not intended to limit the invention in any manner. Those ofskill in the art will readily recognize a variety of non-criticalparameters which can be changed or modified to yield essentially thesame results. The compounds of the Examples have been found to be BETprotein-kinase inhibitors according to at least one assay describedherein.

EXAMPLES Example 1: Characterization of Compound 1 in Xenograft Modelsof JAK2V617F-MPN/AML

Compound and Formulations:

Compound 1 was used in these studies and was manufactured at IncyteCorporation. Ruxolitinib was used in these studies and was manufacturedby Wilmington PharmaTech.

Methods:

Female SCID mice at approximately 5-6 weeks old were subcutaneouslyinoculated with 107 SET-2 cells in matrigel. Dosing began when tumorsize reached approximately 200 mm3. Dosing for these studies wasperformed by oral gavage.

For efficacy studies, measurements of tumors were taken every 2 to 3days using a digital caliper. Tumor volumes were calculated using theformula: Volume=(length×width2)/2 where width was the smaller dimension.Body weights were also monitored.

Animals were sacrificed (1) at end of study, (2) if tumor size reached10% of total body weight, or (3) if 20% of body weight is lost.

Statistical analyses were performed using Prism Graphpad software. Allcomparisons were done using 2-way ANOVA with p<0.05 deemed significant.

Efficacy of Compound 1 in the JAK2 V617F Expressing HEL Xenograft Modelof Erythroleukemia

The purpose of this study was to examine the ability of the BETinhibitor Compound 1 inhibit tumor growth in an MPN-like AML modelexpressing the JAK2 V617F mutation. Female SCID mice were subcutaneouslyinoculated with 107 HEL cells in matrigel. When tumors reachedapproximately 160 mm³, mice (n=10 per group) were dosed orally withCompound 1 for 14 days as follows:

1. BID at either 1 or 3 mg/kg, or vehicle control

2. QD at 3, 10, or 30 mg/kg

As shown in FIG. 1A, administration of Compound 1 inhibited tumor growthin a dose-related manner, and the mean tumor volumes from each of theCompound 1 treatment groups were statistically significantly reducedfrom that of the vehicle group (p<0.02 for each group, 2-way ANOVA). The30 mg/kg QD dose was the most efficacious and resulted in onestatistical tumor regression. The data indicate that dosing Compound 1on a BID regimen could be considered, but is unnecessary, as QD dosingis as effective as BID dosing in this model. All doses were tolerated asdetermined by the lack of body weight loss as shown in FIG. 1B.Efficacy from Combination of Compound 1 and Ruxolitinib in the SET-2Xenograft Model

Female SCID mice were subcutaneously inoculated with 107 SET-2 cells inmatrigel. When tumors reached approximately 175 mm³, mice (n=8 pergroup) were dosed orally for 14 days as follows:

1. Vehicle control, or ruxolitinib at 30 mg/kg BID

2. Compound 1 at 10 mg/kg QD

3. Combination of ruxolitinib and Compound 1

Compound 1 and ruxolitinib were dosed sub-optimally for an optimalread-out of possible combinatorial effects. As seen in FIG. 2A, bothmonotherapies significantly inhibited tumor growth versus vehiclecontrols (p<0.02 for each, 2-way ANOVA). In addition, the combination ofCompound 1 and ruxolitinib gave enhanced and significant (p<0.007 forcombination vs. both single agent arms; 2-way ANOVA) efficacy relativeto expected outcomes based on single agent efficacies. Approximately 15%body weight loss did occur in the combination group (FIG. 2B), but thisnadir was within the limits of tolerability.Efficacy from Combination of Compound 1 and Ruxolitinib in the SET-2Xenograft Model

Female SCID mice were subcutaneously inoculated with 107 SET-2 cells inmatrigel. When tumors reached approximately 150 mm³, mice (n=8 pergroup) were dosed orally for 11 days as follows:

1. Vehicle control, or ruxolitinib at 60 mg/kg BID

2. Compound 1 at 10 mg/kg QD

3. Combination of ruxolitinib and Compound 1

In this study, ruxolitinib was administered at a higher dose than in1.2, resulting in a significant tumor growth inhibition as a monotherapy(FIG. 3A). Compound 1 administration with ruxolitinib did not increasethe tumor growth inhibition seen with single agent ruxolitinib or singleagent Compound 1. All doses were tolerated, although the combination ofCompound 1 and ruxolitinib did result in body weight loss (FIG. 3Bapproximately 12%) within the allowable limit.

The BET inhibitor Compound 1 is efficacious at tolerable doses in twomodels of JAK2V617F AML. Combination of Compound 1 with a sub-optimaldose of the JAK1/2 inhibitor ruxolitinib (30 mg/kg BID) results in asignificant increase in tumor growth inhibition over monotherapies.These data demonstrate the potential for BET inhibitors in combinationwith JAK inhibitors in JAK2V617F driven myeloid neoplasms.

Example 2: Efficacy of Compound 1 in Combination with Ruxolitinib in aMouse Model of MPLW515L-Driven MPN

Compound and Formulations:

Compound 1 and ruxolitinib were used in these studies and weremanufactured by Wilmington PharmaTech.

Methods:

At one week prior to bone marrow transplant, eight week old femaleBalb/c mice (Charles River Laboratories) were injected onceintraperitoneally with 150 mg/kg 5-fluorouracil. Five days later, thesemice were sacrificed, and bone marrow was harvested by aspiration offemurs. Red blood cells were lysed using BD Pharma Lyse buffer (BDBiosciences), then washed with PBS. The remaining marrow was plated in10% FBS/RPMI overnight in standard cell culture incubation. Thefollowing day, the bone marrow was infected with ecotropic retrovirus(MSCV puromycin backbone) expressing the MPLW515L gene by adding 1 mlvirus to 106 cells, then centrifugation at 1800 rpm, 37° C., 90 minutesin 6 well plates. Following centrifugation, infected cells were kept inthe incubator until the following day when they were injected intorecipients.

Bone marrow of recipient mice was ablated in eight week old Balb/c miceusing 50 mg/kg 5-fluorouracil five days prior to transplantation withBalb/c bone marrow expressing MPLW515L via retroviral infection(approximately 2×105 cells per recipient mouse). Blood counts wereperformed 7 days after transplant and 4 cohorts (n=10 per cohort) withequal mean platelet number were generated for treatment. Dosingcommenced on day 8 post implant and continued for 13 days. At day 14post implant, mice were sacrificed, blood collected by cardiac puncture,and spleens weighed as a surrogate for disease burden.

Blood was collected via by orbital sinus once weekly, and complete bloodcounts (CBCs) were determined by hematology instrumentation (Abaxis,model HM5).

Statistical analyses were performed using Prism Graphpad software. Allcomparisons were done using unpaired t tests with p<0.05 deemedsignificant.

Efficacy of Compound 1 in Combination with Ruxolitinib in the MPLW515LModel of Mouse MPN

Female Balb/c mice were transplanted with MPLW515L expressing bonemarrow in order to assess the ability of the BET inhibitor Compound 1 toimprove upon the activity of ruxolitinib alone in a preclinical MPNmodel. On day 7 post transplant, mice were randomized into cohorts fordosing based on platelet levels, as determined by CBC. The following dayoral dosing commenced, with mice receiving Compound 1 at 10 mg/kg QD,ruxolitinib at 60 mg/kg BID, the combination of Compound 1 andruxolitinib, or vehicle control. Dosing proceeded for 14 days, afterwhich mice were bled for CBC, and spleens were harvested and weighed asa surrogate for disease burden. Two vehicle treated mice died before endof study on day 14 post dosing, one on day 12 of dosing and one on day13 post dosing.

While each single agent was able to reduce spleen size, the combinationgave the most efficacy. Mice treated with the Compound 1/ruxolitinibcombination had spleens that were significantly smaller than those ofeither single agent alone (FIG. 4A). Ruxolitinib had a more profoundeffect on white blood cell count reduction than Compound 1, and therewas little difference between WBC counts from ruxolitinib treated miceand mice receiving combination therapy (FIG. 4B).

Statistics were determined by unpaired t tests performed using PrismGraphpad software (**p<0.01; ****p<0.0001).

At the doses chosen for this study, both Compound 1 and ruxolitinib wereable to slow the expansion of white blood cells and suppresssplenomegaly brought on by MPLW515L expressing bone marrow. Combinationof Compound 1 with ruxolitinib resulted in a significant decrease indisease burden over either monotherapy as measured by spleen weight.These data demonstrate the potential for BET inhibitors in combinationwith JAK inhibitors in myeloproliferative neoplasms.

Example 3: Statistical Demonstration of Synergy Between Compound 1 andRuxolitinib

The synergy between Compound 1 and ruxolitinib was assessed using theChou-Talalay equation (see Chou, “Drug Combination Studies and TheirSynergy Quantification Using the Chou-Talalay Method” Cancer Res; 70(2)Jan. 15, 2010, incorporated by reference herein). Synergy depicts agreater effect with the combination than what would be expected based onhow the individual compounds are working. In Table 1, there are data forthe two experiments described herein. The first data set listed is theMPLW515L model, spleen weights are used as disease progression markers(i.e., bigger spleen=worse disease). MPL SPL depicts the sizes of thespleens in grams from the MPLW515L murine MPN model. Fa shows thefraction affected which is the percentage change in comparison to thevehicle. Expected row shows the expected effect of the combination basedon the Chou-Talalay synergy index. Since the number is smaller than theeffect in the fraction affected, this demonstrates a larger effect thanexpected from combining Compound 1 and ruxolitinib demonstrating thatsynergy is occurring between the two compounds giving an unexpectedeffect. The second experiment is SET-2 which is the JAK2V617F expressingcell line used in a xenograft tumor model. Fa for SET-2 are the tumorgrowth inhibition levels for each dose used and the combination. Againthe expected value is smaller than the actual effect demonstrating anunexpected synergy. In both experiments, the Chou-Talalay equation gaveus a value that was smaller than the effect seen with the combinationsof ruxolitinib plus Compound 1, this is interpreted as the combinationbeing synergistic in each experiment.

TABLE 1 Ruxolitinib Compound 1 Vehicle 60 mg/kg BID 10 mg/kg QDCombination MPL SPL 0.812975 0.3367 0.3611 0.0965 Fa 0.5858421230.5558289 0.881300163 Expected 0.81604304 SET-2 Fa 0.44 0.24 0.71Expected 0.5744

Example 4: Clinical Study Protocol of Compound 1 as Monotherapy inParticipants with Myelofibrosis

Study Design

This is a Phase 1, open-label, 2-part study of Compound 1 as monotherapyin participants with relapsed or refractory myelofibrosis (for furtherdetails see “Safety and Tolerability Study of INCB057643 in ParticipantsWith Myelofibrosis”, ClinicalTrials.gov Identifier: NCT04279847, whichis incorporated herein by reference in its entirety). INCB057643 is alsoreferred to as Compound 1 in the present disclosure. Participantsinclude those who have received at least 1 line of prior therapyincluding ruxolitinib and have no further available therapy known toprovide clinical benefit and with a risk category of intermediate-2 orhigh according to DIPSS. Participants will receive 4 mg QD of Compound 1on a continuous basis.

The study will be conducted in 2 parts. Part 1 will evaluate initialsafety and tolerability of 4 mg QD of Compound 1 in participants withrelapsed or refractory myelofibrosis. Study drug will beself-administered once daily every day for 28 days, which is 1 cycle.Participants will continue taking study drug as long as benefit isderived and discontinuation criteria is not met.

If the starting dose of 4 mg QD is deemed tolerable in Part 1, thestarting dose in Part 2 will be 4 mg QD; if not, the starting dose inPart 2 will be 2 mg QD. Part 2 will administer Compound 1 as monotherapyas well.

Throughout the whole study, in both Part 1 and Part 2, AEs will becontinuously monitored. If the cumulative incidence of Grade 3 or 4drug-related AEs occurs in >40% of participants, the study will bestopped. Likewise, if there is more than 1 fatal drug-related event, thestudy will be stopped. The study may only be restarted after discussionwith the FDA.

Pharmacokinetics of Compound 1 as Monotherapy

The clinical PK of Compound 1 in participants with advanced malignancies(solid and hematologic malignancies) were evaluated in a Phase 1/2,open-label, dose-escalation/dose-expansion, safety and tolerabilitystudy. Participants received continuous QD doses of Compound 1. As ofthe data cutoff date (23 Sep. 2019), 25, 101, and 8 participants whoreceived 8 mg, 12 mg, and 16 mg, respectively, across Part 1 and Part 2(Compound 1 treated as monotherapy) have been assessed for PK. The PKparameters of parent Compound 1 are summarized in Table 2 and Table 3(see FIG. 5 ). Table 2 presents a summary of Compound 1 pharmacokineticparameters for Compound 1 as monotherapy (part 1 and 2) at cycle 1day 1. Table 3 presents a summary of Compound 1 pharmacokineticparameters for Compound 1 as monotherapy (part 1 and 2) at steady-state(cycle 1 day 8).

TABLE 2 Partic- Analyte Dose ipant (s) C_(max) (nM) T_(max) (h)AUC_(last) (h*nM) Com-  8 mg N = 12 201 ± 83.6 2.00 2310 ± 1120 pound 1188 (38.8) (1.00, 24.0) 2090 (50.1) (Parent) 12 mg N = 99 266 ± 94.70.50 2740 ± 1090 249 (39.1) (2.00, 8.00) 2550 (39.5) 16 mg N = 7 343 ±93.2 2.00 3370 ± 1060 329 (34.5) (2.00, 4.00) 3210 (34.3) Note: Valuesare presented in the format of mean ± SD and geometric mean (CV %)except that T_(max) is reported as median (range).

Blood samples for the determination of plasma concentrations of Compound1 in Parts 1 and 2 were collected at predose on Cycle 1 Day 1, Cycle 1Day 2, and Cycle 1 Day 8, and at 0.5, 1, 2, 4, 6, and 8 hours on Cycle 1Day 1 and Cycle 1 Day 8. The plasma samples of Compound 1 were assayedby a validated LC-MS/MS method.

With multiple-dose administration in the fasted state, Compound 1 plasmaconcentrations attained the peak values (C_(max)) typically at 1-2 hours(median T_(max)) postdose, and subsequently exhibited a monophasicdecay, with a steady-state geometric mean t_(1/2) of approximately 10hours that was not dose-dependent. The steady-state was predicted to beachieved after 2 days with once daily dosing, based on an effectivehalf-life of 10 hours. There is minimal accumulation of Compound 1exposure (<10%) by comparing steady-state AUC₀₋₂₄ to first dose AUC₀₋₂₄.Within the dose range of 8 to 16 mg QD, increases in the Compound 1steady-state C_(max) and AUC₀₋₂₄ were proportional to dose, that is,Compound 1 exhibited approximately linear pharmacokinetics over the doserange studied. ANOVA of dose normalized PK parameters (C_(max,ss) andAUC_(ss,0-24)) using overall test or pairwise comparisons (see Table 2)demonstrated that dose normalized C_(max,ss) or AUC_(ss,0-24) was notstatistically significantly different (P>0.05) between or across thedoses. Compound 1 exhibited a low steady-state oral clearance withgeometric mean CL_(ss)/F of 9.92-10.7 L/h and moderate volume ofdistribution with geometric mean V_(z)/F of 138-197 L. At a dose of 12mg QD, the geometric mean (CV %) of t_(1/2), steady-state C_(max), andAUC₀₋₂₄ were 9.17 h (49.7%), 272 nM (40.9%), and 2740 h*nM (43.2%),respectively.

The estimated steady-state AUC₀₋₂₄ of Compound 1 is 457 h*nM and 913h*nM for 2 mg QD and 4 mg QD, respectively, based on linear PKextrapolation from 12 mg QD geometric mean AUC_(ss,0-24). The simulatedPK data of Compound 1 at 4 mg QD are presented in FIG. 7 .

Pharmacodynamics for the Treatment of Compound 1 as Monotherapy

Pharmacodynamic analysis was performed using an ex vivo assay measuringprotein levels of cMyc, a BRD4 target gene, in KMS12BM cells, which werespiked into participant plasma samples collected at predose and variouspostdose timepoints. In preliminary PD analysis after 8 mg, 12 mg, and16 mg of oral administration, Compound 1 demonstrated inhibition oftotal cMyc protein expression with maximal inhibition occurring between1 and 4 hours. The average cMyc inhibition at steady state (Day 8, frompredose to 8 hours) was 30%, 44%, and 65% for the dose of 8 mg (n=11),12 mg (n=51), and 16 mg (n=7), respectively. Maximum individual peakinhibition of total cMyc protein expression ranged from 16% to 77% at 8mg, 20% to 92% at 12 mg, and 61% to 97% at 16 mg dose level. Inhibitionof cMyc was reduced to <10% at trough (Cycle 1 Day 8 predose) in the8-mg and 12-mg cohorts and ˜30% in the 16-mg cohort. A composite PK PDcurve was plotted for 70 participants, and an IC50 value of 202.4 nM wasdetermined by nonlinear regression curve fitting (see FIG. 6 ). The exvivo IC50 coverage over steady-state concentrations of Compound 1 areshown in FIG. 7 .

Relationship of Compound 1 Steady-State AUC and Treatment-EmergentAdverse Events

The selected clinical safety endpoints as deemed appropriate (eg,frequently occurring TEAEs (treatment-emergent adverse events), incidentrate >20% for all AEs (adverse events) and clinically notable adverseevents) were used to evaluate exposure-safety relationships. The safetydata from a total of 106 participants (including both solid tumors andhematologic malignancies) receiving monotherapy treatment of Compound 1(n=10 at 8 mg, n=89 at 12 mg, and n=8 at 16 mg) in Parts 1 and 2 wereused for analysis. Frequently occurring TEAEs (all grades and causality)in participants who received at least 1 dose of study drug includednausea (47.9%), fatigue (45.5%), decreased appetite (35.5%),thrombocytopenia (28.9%), vomiting (28.1%), anemia (27.3%), diarrhea(27.3%), constipation (20.7%), and dysgeusia (19.8%) as well asclinically notable AEs including hyperglycemia (17.4%), epistaxis(9.9%), INR increased (8.5%), and gastrointestinal hemorrhage (1.9%).

No statistically significant correlation was identified for steady-stateCompound 1 AUC and any TEAEs or clinically notable AEs evaluated in thisanalysis, except hyperglycemia (p-value<0.05). FIG. 8 showsmodel-predicted versus observed relationship of Compound 1 steady stateAUC and probability of hyperglycemia. The predicted probability ofhyperglycemia incidence at dose of 2 mg and 4 mg is 8.0% and 9.2%,respectively, based on AUC and hyperglycemia relationship (Note: Opensquares represent first (893-2035 h*nM), second (2074-2605 nM), third(2622-3564 nM), and fourth (3606-9788 nM) quartiles of Compound 1AUC_(ss,0-24).). Lack of correlation between Compound 1 AUC_(ss,0-24)and TEAEs could be due to the narrow dose range (8, 12, or 16 mg)explored in this study and small sample sizes in the 8-mg and 16-mggroups of the study (the majority of participants received 12 mg).

Table 4 lists the estimated parameters for Compound 1 AUC and Grade 3INR (international normalized ratio) increases and Grade 4gastrointestinal hemorrhage. Two Grade 3 events of INR occurred inparticipants concomitantly taking warfarin. The steady state AUC ofthese 2 participants were 6990 h*nM and 9770 h*nM receiving Compound 112 mg QD and 16 mg QD, respectively, and the AUCs were much higher thanthe geometric mean steady-state AUC in 12 mg QD (2740 h*nM) and 16 mg QD(3610 h*nM), respectively.

Two Grade 4 events of gastrointestinal hemorrhage were observed in 2participants receiving 12 mg QD dose, and the steady-state AUC of these2 participants were 3470 h*nM and 3610 h*nM, respectively, and the AUCswere slightly higher than geometric mean steady-state AUC in 12 mg QD.

TABLE 4 Participants Adverse Event Grade Dose (mg) AUC_(ss,0-24) (h*nM) 1011 Gastrointestinal 4 12 3470 hemorrhage  9002 Gastrointestinal 4 122150 hemorrhage 12005 INR increased 3 12 6990 16001 INR increased 3 169790

Example 5: Efficacy of Compound 1 in Combination with Ruxolitinib inParticipants with Myelofibrosis

Preliminary efficacy (spleen length and/or volume reduction) wasobserved in 2 of 3 myelofibrosis participants treated in the studyreferenced herein (for further details see “Safety and TolerabilityStudy of INCB057643 in Participants With Myelofibrosis”,ClinicalTrials.gov Identifier: NCT04279847, which is incorporated hereinby reference in its entirety). All three participants were administeredruxolitinib for 6 months prior. A first participant and a secondparticipant had relapsed myelofibrosis which was treated with Compound 1monotherapy. A third participant was being treated with single-agentruxolitinib but having suboptimal responses. In reaction to thesuboptimal responses, the third participant was then treated withCompound 1 in combination with ruxolitinib.

The first participant received Compound 1 monotherapy 12 mg QD and had aspleen length reduction of 92.5% by spleen palpation (nadir compared tothe baseline). The second participant with relapsed myelofibrosisreceived Compound 1 monotherapy 8 mg QD. The second participant on thelower dose of Compound 1 resulted in only disease progression, and thesecond participant stayed on study for 24 days. The third participantreceived Compound 1 8 mg QD in combination with ruxolitinib andexperienced a 77% spleen length reduction by spleen palpation and a 44%spleen volume reduction by imaging (nadir compared to the baseline). Toput the positive results of the first and third participants in context,when a ruxolitinib monotherapy was employed in clinical trials about 35%reduction in spleen size was observed (for further details seeVerstovsek S., Morgan G. “Results of COMFORT-I, a randomizeddouble-blind phase III trial of JAK 1/2 inhibitor INCB18424 (424) vsplacebo (PB) for patients with myelofibrosis (MF)”, Abstract #6500. 2011American Society of Clinical Oncology Annual Meeting, which isincorporated herein by reference in its entirety).

Further preliminary efficacy of a BET inhibitor (CPI-0610) inmyelofibrosis participants was also reported in the MANIFEST study(NCT02158858; Hoffman et al 2019, Mascarenhas et al 2019, which isincorporated herein by reference in its entirety). The enrolledmyelofibrosis participants were either refractory, intolerant, orineligible for ruxolitinib (in the CPI-0610 monotherapy cohort), or werereceiving ruxolitinib but experiencing suboptimal response ormyelofibrosis progression (in the CPI-0610 and ruxolitinib combinationcohort). The participants experienced best spleen volume reduction whichranged from 6%-44% in 10 evaluable participants (monotherapy andcombination combined).

Various modifications of the invention, in addition to those describedherein, will be apparent to those skilled in the art from the foregoingdescription. Such modifications are also intended to fall within thescope of the appended claims. Each reference, including withoutlimitation all patent, patent applications, and publications, cited inthe present application is incorporated herein by reference in itsentirety.

What is claimed is:
 1. A method of treating a myeloproliferativeneoplasm in a patient in need thereof, comprising administering to saidpatient2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3 (4H)-one, or a pharmaceutically acceptablesalt thereof; and ruxolitinib, or a pharmaceutically acceptable saltthereof.
 2. The method of claim 1, wherein the ruxolitinib, or apharmaceutically acceptable salt thereof, is ruxolitinib phosphate. 3.The method of claim 1, wherein2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-oneis a solid form.
 4. The method of claim 3, wherein the solid form is ananhydrate.
 5. The method of claim 3, wherein the solid form is Form I.6. The method of claim 5, wherein Form I has one or more characteristicXRPD peaks, in terms of 2-theta, selected from about 8.7, about 9.8,about 11.6, about 12.7, about 14.7, about 15.7, about 20.0, about 21.4,about 23.3, and about 27.1 degrees.
 7. The method of claim 3, the solidform having Form II.
 8. The method of claim 7, wherein Form II has oneor more characteristic XRPD peaks, in terms of 2-theta, selected fromabout 6.7, about 9.5, about 10.5, about 14.8, about 16.2, about 17.0,about 18.8, and about 19.3 degrees.
 9. The method of claim 1, whereinthe dose of ruxolitinib, or a pharmaceutically acceptable salt thereof,is about 5 mg/day to about 60 mg/day.
 10. The method of claim 1, whereinthe dose of ruxolitinib, or a pharmaceutically acceptable salt thereof,is about 2.5 mg BID to about 30 mg BID.
 11. The method of claim 1,wherein the dose of2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, is about 2 mg/day toabout 20 mg/day.
 12. The method of claim 1, wherein the dose of2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, is about 2 mg/day toabout 18 mg/day.
 13. The method of claim 1, wherein the dose of2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, is about 2 mg/day toabout 12 mg/day.
 14. The method of claim 1, wherein the dose of2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, is about 4 mg/day toabout 8 mg/day.
 15. The method of claim 1, wherein the dose of2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, is about 2, about 4,about 6, about 8, about 10, about 12, about 14, about 16, about 18, orabout 20 mg/day on a free base basis.
 16. The method of claim 1, whereinthe dose of2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, is about 2 mg/day on afree base basis.
 17. The method of claim 1, wherein the dose of2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, is about 4 mg/day on afree base basis.
 18. The method of claim 1, wherein the dose of2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, is about 6 mg/day on afree base basis.
 19. The method of claim 1, wherein the dose of2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, is about 8 mg/day on afree base basis.
 20. The method of claim 1, wherein the dose of2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, is about 10 mg/day on afree base basis.
 21. The method of claim 1, wherein the dose of2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one,or a pharmaceutically acceptable salt thereof, is about 12 mg/day on afree base basis.
 22. The method of claim 1, wherein themyeloproliferative neoplasm is selected from polycythemia vera (PV),essential thrombocythemia (ET), primary myelofibrosis, chronicmyelogenous leukemia (CML), chronic myelomonocytic leukemia (CMML),hypereosinophilic syndrome (RES), systemic mast cell disease (SMCD),chronic neutrophilic leukemia (CNL), and chronic eosinophilic leukemia.23. The method of claim 22, wherein the myeloproliferative neoplasm ispolycythemia vera (PV).
 24. The method of claim 22, wherein themyeloproliferative neoplasm is essential thrombocythemia (ET).
 25. Themethod of claim 22, wherein the myeloproliferative neoplasm is primarymyelofibrosis.
 26. The method of claim 1, wherein ruxolitinib, orpharmaceutically acceptable salt thereof, and2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one, or a pharmaceutically acceptablesalt thereof, are administered orally.
 27. The method of claim 1,wherein ruxolitinib, or pharmaceutically acceptable salt thereof, and2,2,4-trimethyl-8-(6-methyl-7-oxo-6,7-dihydro-1H-pyrrolo[2,3-c]pyridin-4-yl)-6-(methylsulfonyl)-2H-1,4-benzoxazin-3(4H)-one, or a pharmaceutically acceptablesalt thereof, are administered simultaneously or sequentially.